Qingxuan Jiangya Decoction Mitigates Renal Fibrosis In Spontaneously Hypertensive Rats By Regulating Transforming Growth Factor-?1/Smad Signaling Pathway

Objective:To examine the therapeutic efficacy of QXJYD against elevated blood pressure using the SHR model,as well as the mechanisms behind its anti-hypertensive activity and protection against renal fibrosis.To further illuminate the underlying mechanisms of QXJYD in protecting against hypertension,provides a theoretical basis for the widespread use of QXJYD in the prevention and treatment of hypertension.Methods:1.Blood pressures of 20 male Spontaneously Hypertensive(SHR)rats and 10 normotensive control Wistar Kyoto(WKY)rats were measured once per week,with the tail-cuff plethysmograph method using CODATM non-invasive blood pressure system.All rats were acclimatized for one week prior to experiments.After 8 weeks,the rat's kidneys were eviscerated.2.Examined the histological changes in the kidneys of rats using Harris Hematoxylin and Eosin(H&E)staining.3.The degree of collagenous fiber formation in the kidneys of rats were measured using Masson's trichrome staining.The areas of Masson's trichrome-positive was analysed by Semi-quantitative.4.Immunohistochemical staining to analyze the expression of NOX2,NOX4,TNF-?;IL-6 and Collagen ?,Collagen ?,Collagen ? in the kidney.5.Real-time PCR detected the mRNA expression of TGF-?1 Smad3,MMP-2,MMP-9,Collagen ?,Collagen ?,Collagen IV.6.Western blot analysed the protein expression and activation of TGF-?1,Smad2/3,Smad4,MMP-2,MMP-9 in TGF-?/Smad signaling pathway.Results:1.According to results of blood pressure,after treatment with QXJYD for six weeks has significantly attenuated the elevation of SBP,DBP,MAP in rats.Moreover,administration of QXJYD did not result in body weight changes in experimental rats.2.H&E staining showed that kidney of SHRs had significantly enlarged glomerulus and atrophy compared to the normotensive WKY rats,and exhibited significant injuries to the tubulointerstitium.However,the injures to the glomerulus in SHRs was robustly mitigated by QXJYD treatment.3.Masson's trichrome staining founded that there were extensive areas of collagen fiber deposition within the interstitial space in SHRs,which was reversed by QXJYD treatment.4.Immunohistochemical staining suggested the high expressions of NOX2,NOX4,TNF-a,IL-6 and Collagen ?,Collagen ?,Collagen ? in SHRs,which was robustly attenuated following QXJYD treatment.5.Real-time PCR detected the mRNA levels of TGF-?1,Smad3,MMP-2,MMP-9 and Collagen ?,Collagen ?,Collagen ? were all promoted in SHRs group,which was reversed following QXJYD treatment.6.Western blot showed that the protein expression of TGF-?1,Smad2/3,Smad4,MMP-2,MMP-9 were all increased in SHRs group,which was also attenuated following QXJYD treatment.Conclusion:1.QXJYD could significantly attenuate the elevation of blood pressure in an SHR model,but had no effect on body weight changes.2.QXJYD could decreased the level of NOX2,NOX4 and TNF-a,IL-6 in SHR model.Meanwhile attenuate the content of collagen caused by SHR model.3.QXJYD treatment also mitigated the content of Collagen ?,Collagen ?,Collagen? and the expression of MMP-2,MMP-9 in SHR group.4.QXJYD treatment also mitigated the renal fibrosis via regulated the TGF-?1/Smad signaling pathway.